1.Effect of chloroform extract of Zingiber mioga Roscoe on human dermal microvascular endothelial cell expression of cell adhesion molecules and adhesivity to lymphocytes
Zhongzhu LI ; Qiguo ZHANG ; Weihua LI ; Songlin SHI ; Qifu LI
Chinese Journal of Dermatology 2013;46(10):726-730
Objective To investigate the effect of chloroform extract of Zingiber mioga Roscoe on human dermal microvascular endothelial cell (HDMEC) expression of cell adhesion molecules and adhesivity to lymphocytes.Methods HDMECs were isolated from human foreskin tissue and subjected to subculture.After several passages,some HDMECs were treated with chloroform extract from Zingiber mioga Roscoe (CFMG) of 200 mg/L for 30 minutes before or after 24-hour stimulation with tumor necrosis factor (TNF)-α or interleukin (IL)-1α.Subsequently,enzyme-linked immunosorbent assay was performed to determine the expression levels of intercellular adhesion molecule-1 (ICAM-1),vascular cell adhesion molecule-1 (VCAM-1) and E-selectin on HDMECs.Both T lymphocytes isolated from venous blood of healthy adults and Ramos Burkitt's lymphoma B cells were labelled with chromium-51 and incubated with the HDMECs for four hours followed by the determination of radiation intensity of lymphocytes adhering to HDMECs using a gamma-counter.Results Compared with 0.2% dimethyl sulfoxide (DMSO),CFMG slightly down-regulated the expression of ICAM-1,VCAM-1 and E-selectin on resting HDMECs (all P > 0.05).In comparison with culture medium,TNF-α enhanced the expressions of ICAM-1,VCAM-1 and E-selectin significantly (all P < 0.01),and IL-1α elevated the expressions of ICAM-1 and E-selectin markedly(both P < 0.01) as well as the expression of VCAM-1 slightly (P > 0.05).The upregulating effects of TNF-α and IL-1α on the expressions of adhesion molecules were notably suppressed by the CFMG treatment prior to the stimulation with TNF-α and IL-1α(all P < 0.01),but not affected by that after the stimulation (all P > 0.05).The adhesivity of HDMECs to T lymphocytes and Ramos cells was slightly decreased by CFMG treatment compared with 0.2% DMSO (both P > 0.05),but was markedly increased by TNF-α and IL-1α compared with the culture medium (both P < 0.01).Conclusions CFMG may play an antiinflammatory role via blocking the up-regulating effect of pre-inflammatory cytokines such as TNF-α and IL-1α on the expression of adherence molecules on HDMECs and,hence,inhibiting inflammatory cell infiltration in tissue.
2.Effect of ascending blood formula on immune function and bone marrow suppression in patients with colon cancer chemotherapy
Meifang YAO ; Fengyun LI ; Zhi LIU ; Zhongzhu FU ; Huarong LIU
Chongqing Medicine 2016;45(27):3799-3801
Objective To explore the influence of the old Chinese national match the experience of haplostele liters of blood prescription of chemotherapy for colorectal cancer patients with immune function and bone marrow suppression .Methods 94 cases of metastatic nodes in patients with rectal cancer treated in our hospital from February 2011 to June 2015 were selected as the re‐search object ,and were randomly divided into observation group and control group according to different regimen in the treatment , with 47 cases in each group .Patients in control group was treated with conventional XELON chemotherapy ,and patients in the ob‐servation group recieved rise blood prescription treatment based on the chemotherapy in the control group .The immune function and bone marrow suppression situation in both groups were observed after two treatment cycles .Results The stable rate of the obser‐vation group was 42 .55% ,significantly higher than that of the control group (P<0 .05);the CD4+ /,CD8+ CD8+ and NK cells were significantly higher than those in the control group (P<0 .05) ,while those of the control group was significantly higher than that in control group (P>0 .05);the CD4+ /,CD8+ CD8+ and CD4+ /cells were significantly higher than those in control group (P<0 .05);The incidence rate of KPS in the observation group and control group was 70 .21% ,70 .21% ,17 .02% ,14 .89% , 87 .23% ,25 .53% and 23 .40% respectively .The KPS score of the observation group and the control group were significantly higher than that in control group (P<0 .05) ,and the G‐CSF in the observation group was significantly higher than that in control group (P<0 .05) .Conclusion The treatment of advanced metastatic colorectal cancer patients in the course of chemotherapy to give rise to blood formula can effectively improve the patient′s immune function ,reduce the bone marrow suppression effect ,improve the quality of life ,and it is worthy of clinical application .
3.Overexpression of HIV-1 P17 protein in Escherichia coli and its purification
Zhongzhu LI ; Ningyi JIN ; Hongwei WANG ; Zhiru GUO ; Ping LI ; Hangyi YANG ; Zhen YIN
Journal of Jilin University(Medicine Edition) 2001;27(1):19-21
Objective:To highly express HIV-1p17 in E.coli and purify the protein.Methods:①Recombinant plasmid was constructed by inserting HIV-1p17 gene amplified by PCR into plasmid vector,pET28c;②The recombinant plasmid was expressed in BL21,BL21(DE3),BL21(DE3) plysS and HMS174(DE3) of E.coli separately;③The target protein were purified with Ni-NTA resin;④The purified protein was detected by western blot and ELISA.Results:The expression of the P17 protein in BL21(DE3) represented up to 32% of total protein in E.coli,which was the most amounts compared with other kinds of E.coli.The purity of the purified protein reached 95%.The purified protein was recognized by HIV-1P17McAb as well as by HIV-1 positive serum.Conclusion:The recombinant plasmid is highly expressed in BL21(DE3) of E.coli that can be proceeded to the immunocompetence and the bioactivity research.The method of Ni-NTA resin is simple with low protein losing and high purity.And the purified p17 can be employed in early detection of HIV-1 infection and prediction of the clinical progression.
4.Tumor antigen-pulsed dendritic cell vaccine for treatment of patients with advanced malignant tumor:a clinical observation
Liming ZHU ; Yiping ZHANG ; Xiaojia WANG ; Haijun ZHONG ; Yun FAN ; Xinmin YU ; Qinghua DENG ; Xiangming KONG ; Jieer YING ; Tie LI ; Guangyuan LOU ; Zhibing WU ; Zhongzhu TANG ; Shengling MA
Chinese Journal of Cancer Biotherapy 2006;0(06):-
Objective: To observe the safety and clinical efficacy of tumor antigen-pulsed dendritic cell(DC) vaccine in treatment of advanced malignant tumor.Methods: Ninety-one patients with non-small cell lung cancer,colon and rectal cancer,melanoma,renal carcinoma,breast cancer and other malignant tumors were enrolled in this study.All patients met the selecting standard and signed informed consent.Human dendritic cells were obtained from peripheral blood monocytes by culturing them with granulocyte macrophage-colony stimulating factor and interleukin-4.DC vaccine was prepared from tumor antigen pulsed immature dendritic cells in vitro.Patients received the vaccine therapy once every week and one cycle was defined as once every week for 3 weeks.Results: All the patients received 96 cycles of DC vaccine treatment.Symptoms of toxicity included fever,shivering,aching pain of muscle,asthenia,itching,stifle and transient fatigue;most of the symptoms automatically recovered.Clinical efficacy of the treatment was evaluated in 76 patients.Thirty-one of the 76 patients were stable after treatment and 45 were in progressive situation,with the clinical benefiting rate being 40.8%.Eighty-five patients were followed up.The median time for progression was 2.6 months;the overall survival time was 0.9-30.6 months;and the median survival period was 4.5 months,with the one year survival rate being 9.2%.Conclusion: The results suggest that the DC vaccine therapy is well tolerated in treating patients with advanced malignant tumors and has satisfactory clinical benefit;the clinical value of DC vaccine therapy needs to be further observed.